Method for identifying autoimmune disease, method for detecting anti-Reg protein autoantibody and diagnostics for autoimmune disease

Abstract

A method for judging an autoimmune disease by detecting the existence of an anti-Reg protein autoantibody in a specimen; and a method for judging insulin-dependent or non-insulin-dependent diabetes mellitus.A method for detecting an anti-Reg protein autoantibody by bringing into a specimen into contact with an antigen component and detecting the formation of an immune complex.A reagent for diagnosing autoimmune disease which contain an antigen component capable of binding specifically to the anti-Reg protein autoantibody; and a reagent for diagnosing insulin-dependent or non-insulin-dependent diabetes mellitus.

Description

TECHNICAL FIELD[0001]This application is the U.S. National Phase under 35 U.S.C. §371 of International Application PCT / JP00 / 02245, filed Apr. 6, 2000, which claims priority to Japanese Patent Application No. 11-99963, filed Apr. 7, 1999. The International Application was not published under PCT Article 21(2) in English.[0002]The present invention relates to a method for identifying autoimmune diseases and diabetes mellitus (including insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, etc.). The present invention also relates to a method for detecting an anti-Reg protein autoantibody, and a reagent diagnosing for autoimmune diseases and diabetes mellitus (including insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus, etc.).BACKGROUND ART[0003]Autoimmune diseases are diseases caused by an abnormal immune system in which the immune system does not work normally to produce humoral or cellular immune response to self-cells or self-tissu...

AI Technical Summary

Benefits of technology

[0051]As for the antigen components used in the method for detection of an anti-Reg protein autoantibody of the present invention, it is possible to use the fragmentated Reg proteins by means of well-known enzymatic procedures or genetic recombination technique or the partially modified Reg proteins by means of well-known enzymatic procedures or genetic recombination technique, in view of their sensitivity and specificity in detection of the anti-Reg protein autoantibody as well as the stability of antigenic activity during preservation as a diagnostic reagent for a long period.

[0079]In addition, purification from the culture broth and from the incubated transformant as for a culture may be combined to improve yield of the objective Reg protein. Crushing of the transformant may be carried out, for example, by a method for physically crushing the transformant. For example, the transformant is suspended in a buffer, to which ultrasonic wave is irradiated, or the transformant blended with quartz sand is suspended in a buffer. On the other hand, dissolution of the transformant may be achieved, for example, by a method for dissolving the cell wall of the transformant with an enzyme and a surfactant. When the host transformant is Escherichia coli, lysozyme as enzyme and sodium dodecyl-sulfate (hereinafter abbreviated to SDS) as surfactant may be used.

[0094]Electrophoresis of Reg proteins may be achieved, for example, by a variety of known electrophoreses such as SDS polyacrylamide electrophoresis, native polyacrylamide gel electrophoresis, gel isoelectric focusing, etc. Among them, SDS polyacrylamide electrophoresis is preferred since the proteins can be separated based on their molecular weight and the transcription is efficient.

Problems solved by technology

Thus, though the positive rate of autoantibody is high in IDDM, it is low in NIDDM, and there is no report on an effective autoantibody as a diagnostic marker for NIDDM.

However, so far there is no report on the existence of autoantibodies against Reg proteins and autoimmune diseases caused thereby.

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